摘要
目的:采用RNA干扰技术探讨过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)表达在人胃癌中的作用。方法:根据人PPARγ基因序列设计获得3个小干扰RNA(small interfering nRNA,siRNA)靶位点,并构建了3个针对靶位点的一个载体编码一个短发夹状RNA(shRNA)的质粒表达载体(Y1、Y2和Y3),同时合成了1个阴性对照质粒表达载体(HK),采用阳离子脂质体将质粒转染人胃癌MGC803细胞,FCM法观察转染效率。分别采用RT-PCR和Western印迹法检测PPARγmRNA和蛋白的表达,MTT法检测细胞增殖,相差倒置显微镜、Hoechst33342染色和FCM检测细胞凋亡。结果:RNA干扰(RNA interference,RNAi)沉默PPARγ表达后抑制人胃癌MGC803细胞增殖并诱导其凋亡。结论:PPARγ高表达可能通过促进细胞增殖和抑制细胞凋亡促进人类胃癌的形成,PPARγ有望成为治疗胃癌的新靶点。
Objective： To explore the role of peroxisome proliferator-activated receptor γ （PPARγ） expression in human gastric carcinoma using RNA interference technology. Methods： Three targeted locations were designed and three plasmid vectors （ Y1, Y2 and Y3） expressing short hairpin RNA （shRNA） tartgeting PPAR＇y were constructed according to transcription RNA location of human PPARγ gene sequence and designing principle of small interference RNA （siRNA）. Also a negative control plasmid vector HK was constrncted. They were transfeeted into human gastric carcinoma MGC803 cells mediated by cationic liposome vector; the transfection efficiency of plasmid vector was evaluated by flow cytometry （FCM） ; PPARγ, mRNA and protein expression levels were detected by RT-PCR and Western blotting, respectively. Proliferation of cells was measured by MTT assay and cell apoptosis was detected by contrast phase microscopy, Hoechst 33342 staining, and FCM, respectively. Results： Silencing PPARγ by RNA interference （RNAi） inhibited proliferation and induced apoptosis of human MGC803 ceils. Conclusion：These results suggest that PPARγ over-expression suppressed the carcinogenesis of human gastric cancer through inhibiting proliferation and inducing apoptosis. PPARγ has the potential to become a new target for treatment of gastric carcinoma.
出处
《肿瘤》
CAS
CSCD
北大核心
2008年第10期855-858,868,共5页
Tumor
